Light-based technologies of different wavelengths can inactivate pathogenic microorganisms, but each wavelength has its limitations. This work explores the potential of sequential treatments with different wavelengths for enhancing the disinfection performance of individual treatments by employing various bactericidal mechanisms. The effectiveness, inactivation kinetics, and bactericidal mechanisms of treatments with 222/405, 280/405, and 405 nm alone against Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, Salmonella Typhimurium, and Pseudomonas aeruginosa were evaluated. Inactivation experiments were performed in thin liquid bacterial suspensions that were treated either individually with 48 h of 405-nm light or sequentially with (i) 30 s of 222-nm far-UV-C light, followed by 48 h of 405-nm light, or (ii) 30 s of 280-nm far-UV-C light, followed by 48 h of 405-nm light. Survivors were recovered and enumerated by standard plate counting. All inactivation curves were non-linear and followed the Weibull model (0.99 ≥ R2 ≥ 0.70). Synergistic effects were found for E. coli, L. monocytogenes, and S. Typhimurium, with maximum inactivation level increases of 2.9, 3.3, and 1.1 log CFU after the sequential treatments, respectively. Marginal synergy was found for S. aureus, and an antagonistic effect was found for P. aeruginosa after sequential treatments. Significant differences in reactive oxygen species accumulation were found (P < 0.05) after various treatment combinations, and the performance of sequential treatments was correlated with cellular oxidative damage. The sequential wavelength treatments proposed demonstrate the potential for enhanced disinfection of multiple foodborne pathogens compared with individual wavelength treatments, which can have significant food safety benefits. IMPORTANCE Nonthermal light-based technologies offer a chemical-free method to mitigate microbial contamination in the food and healthcare industries. However, each individual wavelength has different limitations in terms of efficacy and operating conditions, which limits their practical applicability. In this study, bactericidal synergism of sequential treatments with different wavelengths was identified. Pre-treatments with 280 and 222 nm enhanced the disinfection performance of follow-up 405-nm treatments for multiple foodborne pathogens by inducing higher levels of cellular membrane damage and oxidative stress. These findings deliver useful information for light equipment manufacturers, food processors, and healthcare users, who can design and optimize effective light-based systems to realize the full potential of germicidal light technologies. The results from the sequential treatments offer practical solutions to improve the germicidal efficacy of visible light systems, as well as provide inspiration for future hurdle disinfection systems design, with a positive impact on food safety and public health.
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